TGF-beta mediated G1 arrest in a human melanoma cell line lacking p15INK4B: evidence for cooperation between p21Cip1/WAF1 and p27Kip1

We have studied TGF-beta mediated G1 arrest in WM35, an early stage human melanoma cell line. These cells have lost p15INK4B expression through loss of one chromosome 9 and rearrangement of the other. In asynchronously growing WM35, TGF-beta caused reductions in cyclin D1, cyclin A and cdk4 proteins and their associated kinase activities and an increase in both p21Cip1/WAF1 and p27Kip1. These findings were confirmed in cells released from quiescence in the presence of TGF-beta, in which TGF-beta inhibited or delayed the reduction in the cdk inhibitors that normally occurs in late G1. In contrast to observations in other cell types, there was an increased association of both p21Cip1/WAF1 and p27Kip1 with cyclin D1/cdk4 and with cyclin E/cdk2 during TGF-beta mediated arrest of asynchronously growing cells. Upregulation of p21Cip1/WAF1 preceded that of p27Kip1. Furthermore, p21Cip1/WAF1 and p27Kip1 were not present in the same cdk complexes but bound distinct populations of target cdk molecules. Both p21Cip1/WAF1 and p27Kip1 immunoprecipitates from asynchronously growing cells contained active kinase complexes. These KIP-associated kinase activities were reduced in TGF-beta arrested cells. It has been proposed that in TGF-beta arrested epithelial cells, up-regulation of p15INK4B and of p15INK4B binding to cdk4 serves to destabilize the association of p27Kip1 with cyclin D1/cdk4, promoting p27Kip1 binding and inhibition of cyclin E/cdk2. Our findings demonstrate that this is not a universal mechanism of G1 arrest by TGF-beta. In TGF-beta arrested WM35, which lack p15INK4B, the increased p21Cip1/WAF1 may serve a similar function to that of p15INK4B: initiating kinase inhibition and providing an additional mechanism to supplement the effect of p27Kip1 on G1 cyclin/cdks.     

Silicon-substrate microelectrode arrays for parallel recording ofneural activity in peripheral and cranial nerves

A new process for the fabrication of regeneration microelectrode arrays for peripheral and cranial nerve applications is presented. This type of array is implanted between the severed ends of nerves, the axons of which regenerate through via holes in the silicon and are thereafter held fixed with respect to the microelectrodes. The process described is designed for compatibility with industry-standard CMOS or BiCMOS processes (it does not involve high-temperature process steps nor heavily-doped etch-stop layers), and provides a thin membrane for the via holes, surrounded by a thick silicon supporting rim. Many basic questions remain regarding the optimum via hole and microelectrode geometries in terms of both biological and electrical performance of the implants, and therefore passive versions were fabricated as tools for addressing these issues in on-going work. Versions of the devices were implanted in the rat peroneal nerve and in the frog auditory nerve. In both cases, regeneration was verified histologically and it was observed that the regenerated nerves had reorganized into microfascicles containing both myelinated and unmyelinated axons and corresponding to the grid pattern of the via holes. These microelectrode arrays were shown to allow the recording of action potential signals in both the peripheral and cranial nerve settings, from several microelectrodes in parallel     

The cholesterol-lowering properties of a psyllium-based breakfast cereal in hamsters

Hamsters were fed high fiber diets containing cellulose, wheat bran or psyllium. The psyllium was incorporated into high fiber, ready-to-eat (RTE) flakes that were used to formulate the test diet. All the diets contained 0.125% cholesterol. The study was terminated after three weeks. Food intake, weight gain and feed efficiency were not significantly different in the three groups. Serum total and HDL cholesterol levels were reduced significantly by the psyllium diet. Serum triglycerides were 26% lower in the hamsters fed psyllium but because of the large variation the difference did not reach statistical significance. Liver total cholesterol and cholesteryl ester levels were significantly lower in the hamsters fed psyllium. Liver triglycerides were highest in the psyllium-fed hamsters and liver phospholipid levels were similar in the three groups. Liver cholesterol and triglyceride levels were higher in hamsters fed cellulose than in those fed wheat bran. Psyllium formulated into an RTE cereal was effective in reducing serum and liver cholesterol levels in hamsters.     

Ligands with affinity to specific sequences of DNA base pairs. X. Synthesis and binding of netropsin analogs, containing a chelating copper ion peptide, with DNA

An analogue of netropsin has been synthesized consisting of two N-propylpyrrolcarboxamide units linked covalently to a copper-chelating tripeptide Gly-Gly-L-His by means of two and three glycine residues. Binding to DNA and synthetic polynucleotides of netropsin analogue containing three glycine residues between Gly-Gly-L-His tripeptide and the N-end of netropsin analogue (His-Nt) has been studied. It is shown that this netropsin analogue chelates a copper ion with 1:1 stoichiometry, similar to a free Gly-Gly-L-His peptide. It is found that this netropsin analogue occupies 3 to 4 base pairs upon binding to poly(dA).poly(dT) and poly[d(AT)].poly[d(AT)] polymers, irrespective of whether it binds in Cu(2+)-ligated or unligated forms. Binding constants and binding site sizes have been calculated for netropsin analogue complexes with DNA, poly(dA).poly(dT) and poly[d(AT)].poly[d(AT)] polymers at the [Cu2+]/[His-Nt] ratio equal to 0 and 1.0. In the three-component system including His-Nt and Cu(2+)-His-Nt, cooperative effects are recognized which can be explained by heterodimer generation on interaction of His-Nt and Cu(2+)-His-Nt at adjacent binding sites.     

Early detection of ageing in solder joints through laser probe thermal analysis of the peltier effect

We have recently developed an optical contactless method for testing the quality of solder joints during accelerated thermal cycling ageing processes.¹ The method was based upon the measurement of the dynamic thermal behaviour of the joint to short bursts of Joule heating. It has proved to be efficient in revealing the formation of cracks at the lead-solder interface. We present a method to evaluate ageing at a much earlier stage in the cycling process. We have observed in earlier work,¹ that before cracks appear, structural changes occur in the solder at the lead-solder interface. The thermal response of the solder joint is recorded over time to a Peltier heat perturbation produced by flowing a current pulse through the interface where structural changes occur. The key point in this method is to discriminate the Peltier effect from the Joule thermal response because both effects generate heat. The variation of the early Peltier response in the thermal cycling ageing tests is seen as a quantitative signature of the structural changes in the lead-solder interface.     

The Bi2O3-SrO phase diagram

The phase relations in the Bi₂O₃-SrO system have been examined in air at temperatures above 740 °C. Intermediate phases were found to agree with previous work, but the homogeneity ranges and high-temperature phase relationships were in some cases at variance. A modified phase diagram is proposed.     

Image compression using the 2-D wavelet transform

The 2-D orthogonal wavelet transform decomposes images into both spatial and spectrally local coefficients. The transformed coefficients were coded hierarchically and individually quantized in accordance with the local estimated noise sensitivity of the human visual system (HVS). The algorithm can be mapped easily onto VLSI. For the Miss America and Lena monochrome images, the technique gave high to acceptable quality reconstruction at compression ratios of 0.3-0.2 and 0.64-0.43 bits per pixel (bpp), respectively.     

Kalman decomposition for implicit linear systems

The Kalman decomposition for a general class of implicit systems is introduced and studied. The properties of strong reachability, observability, acceptance of all inputs and outer sequences, output-uniqueness, uniqueness, and trajectory observability of the original system are related to the properties of its subsystems obtained via the Kalman decomposition. The problems of transfer relation, regularizability by feedback, output feedback, and output injection are studied by means of this decomposition